Epithelial cell adhesion molecule (EpCAM) is a transmembrane glycoprotein functioned as cell-cell adhesion in epithelia, cell migration, proliferation and differentiation. EpCAM have oncogenic potential through upregulation of c-myc, cyclins A & E after cleavage. Epcam is not only used as diagnostic marker for various cancer, but also a potential target for cancer therapy.
Cell Line Name:
ID8 EPCAM Cell Line
Host Cell Line:
Mouse ID8 cell line
Stable ID8 clone expressing exogenous human EPCAM
One vial of frozen cells (5X106 per vial)
Stable in culture over a minimum of 10 passages
Drug screening and biological assays
70% DMEM+20% FBS+10% DMSO
Split saturated culture 1:4 every 3 days; seed out at about 1-3 × 105 cells/mL
37 °C with 5% CO2
Liquid nitrogen immediately upon receiving
Cell Line Generation
ID8 human EPCAM cell Line was generated using a retroviral vector expressing human EPCAM sequence.
Characterization using PCR sequencing and WB
1. Prewarm culture medium (DMEM + 10% FBS + 2μg/mL Puromycin) in a 37°C water bath.
2. Thaw the frozen vial in a 37°C water bath for 1-2 minutes.
3. Transfer the vial into biosafety cabinet, and wipe the surface with 70% ethanol.
4. Unscrew the top of the vial and transfer the cell suspension gently into a sterile centrifuge tube containing 9.0mL complete culture medium.
5. Spin at ~ 125 × g for 5-7 minutes at room temperature, and discard the supernatant without disturbing the pellet.
6. Resuspend cell pellet with the appropriate volume of complete medium and transfer the cell suspension into a T25 culture flask.
7. Incubate the flask at 37°C, 5% CO2 incubator.
8. Split saturated culture 1:4 every 3 days; seed out at about 1-3 × 105 cells/mL.
1. Prepare the freezing medium (70% DMEM + 20% FBS + 10% DMSO) fresh immediately before use.
2. Keep the freezing medium on ice and label cryovials.
3. Transfer cells to a sterile, conical centrifuge tube, and count the cells.
4. Centrifuge the cells at 250×g for 5 minutes at room temperature and carefully aspirate off the medium.
5. Resuspend the cells at a density of at least 3×106 cells/mL in chilled freezing medium.
6. Aliquot 1 mL of the cell suspension into each cryovial.
7. Freeze cells in the CoolCell freezing container overnight in a -80°C freezer.
8. Transfer vials to liquid nitrogen for long-term storage
1. Litvinov, Sergey; et al. (1994). "Ep-CAM: a human epithelial antigen is a homophilic cell–cell adhesion molecule". The Journal of Cell Biology. 125 (2): 437–46.
2. Maetzel, Dorothea; et al. (2009). "Nuclear signalling by tumour-associated antigen EpCAM". Nature Cell Biology. 11 (2): 162–71.
3. Osta, WA; et al. (2004). "EpCAM is overexpressed in breast cancer and is a potential target for breast cancer gene therapy". Cancer Res. 64 (16): 5818–24.
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